Endophytic bacteria reside within plant hosts without causing disease symptoms. In this study, 853 endophytic strains were isolated from aerial tissues of four agronomic crop species and 27 prairie plant species. We determined several phenotypic properties and found approximately equal numbers of gram-negative and gram-positive isolates. In a greenhouse study, 28 of 86 prairie plant endophytes were found to colonize their original hosts at 42 days postinoculation at levels of 3.5 to 7.7 log 10 CFU/g (fresh weight). More comprehensive colonization studies were conducted with 373 corn and sorghum endophytes. In growth room studies, none of the isolates displayed pathogenicity, and 69 of the strains were recovered from corn or sorghum seedlings at levels of 8.3 log 10 CFU/plant or higher. Host range greenhouse studies demonstrated that 26 of 29 endophytes were recoverable from at least one host other than corn and sorghum at levels of up to 5.8 log 10 CFU/g (fresh weight). Long-range dent corn greenhouse studies and field trials with 17 wild-type strains and 14 antibioticresistant mutants demonstrated bacterial persistence at significant average colonization levels ranging between 3.4 and 6.1 log 10 CFU/g (fresh weight) up to 78 days postinoculation. Three prairie and three agronomic endophytes exhibiting the most promising levels of colonization and an ability to persist were identified as Cellulomonas, Clavibacter, Curtobacterium, and Microbacterium isolates by 16S rRNA gene sequence, fatty acid, and carbon source utilization analyses. This study defines for the first time the endophytic nature of Microbacterium testaceum. These microorganisms may be useful for biocontrol and other applications.
Four distinct viruses with double-stranded DNA are known to replicate in Chlorella-like algae symbiotic with hydras and paramecia. An attempt was made to infect a number of cultured Chlorella strains derived from invertebrate hosts with these viruses. One of the viruses, PBCV-1, replicated in two of the algal strains. Restriction endonuclease analysis of the viral DNA showed that the infectious progeny virus was identical to the input virus; thus, Koch's postulates were fulfilled. Viral infection of the two Chlorella strains has allowed the large-scale production of a eukaryotic algal virus and the development of a plaque assay for the virus.
We previously reported that isolation of symbiotic Chloreila-like algae from the Florida strain ofHydra viridis induced replication of a virus (designated HVCV-1) in the algae. We now report that isolation ofsymbiotic Chlorella-like algae from four other sources of green hydra and one source ofthe protozoan Paramecium bursaria also induced virus synthesis. Algae from one of these hydra contained a virus identical to HVCV-1 (based on its rate of sedimentation, buoyant density, reaction to HVCV-1 antiserum, and DNA restriction fragments) whereas algae from the other three hydra contained. another similar, but distinct, virus (designated HVCV-2). The virus from the paramecium algae (designated PBCV-1) was distinct from both HVCV-1 and HVCV-2. The symbiotic algae in the hydra could also be distinguished ultrastructurally. Chloroplasts of both algae that produced HVCV-1 lacked a pyrenoid whereas chloroplasts of the other three symbiotic algae contained pyrenoids. Since all symbiotic eukaryotic algae we have examined have had virus, a potential viral role in symbiosis is suggested.
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